Molecular marker related to azoospermia as well as detection method and application of molecular marker

A technique for molecular markers and azoospermia, which is applied in the fields of analytical chemistry and clinical medicine, can solve the problems of not being able to fully reflect the overall status of semen and effectively diagnosing male infertility, so as to delay and prevent disease progression, be easy to detect, The effect of improving sensitivity and specificity

Active Publication Date: 2022-04-29
苏州南医大创新中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More importantly, the traditional semen parameter examination cannot fully reflect the whole status of semen
Therefore, male infertility often manifests as no obvious abnormalities in conventional semen parameters, which also makes traditional semen parameters examinations ineffective in diagnosing male infertility.

Method used

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  • Molecular marker related to azoospermia as well as detection method and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1 Research Object Selection and Grouping Basis

[0075] The subjects of this part of the study were first-diagnosed azoospermia cases and normal controls with normal semen parameters from the Affiliated Hospital of Nanjing Medical University and the Reproductive Medicine Center of the Sixth Affiliated Hospital of Sun Yat-sen University. The research content and the informed consent form were approved by the Ethics Committee of Nanjing Medical University and complied with the requirements of relevant laws and regulations. The cases and controls signed the informed consent form after understanding the contents. All the research subjects underwent a complete physical examination and completed a report that included personal basic information, living habits, occupational and environmental exposures, genetic risk factors, sexual function and reproduction. Questionnaires on function, disease history, and physical activity. Specific sample classification criteria a...

Embodiment 2

[0090] Example 2 UPLC-MS metabolomics screening of azoospermia 1,3-dihydroxypropan-2-yl pentadecanoate

[0091] 1. Sample pretreatment

[0092] 1.1. Take 10 μL of semen, add 10 μL of internal standard A, add 10 μL of internal standard B, add 10 μL of internal standard C, add 40 μL of methanol (reagent A), vortex for 30 seconds, and precipitate protein.

[0093] 1.2. Centrifuge at 16000g at 4°C for 15min in a centrifuge, transfer the supernatant to a 1.5mL imported EP tube, and concentrate the supernatant to dryness in a centrifugal concentrator at room temperature.

[0094] 1.3. Reconstitute with 10 μL of ultrapure water (reagent D) and wait for analysis.

[0095] 2. Instrument testing

[0096] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0097] 2.2. Liquid phase conditions:

[0098] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chromatographic column ...

Embodiment 3

[0109] Example 3 Diagnosis of azoospermia by 1,3-dihydroxypropan-2-ylpentadecanoate

[0110] According to the above-mentioned UPLC-Q exactive MS metabolomics method, the inventors detected 1,3-dihydroxypropan-2-ylpentadecanoate in the semen samples of 62 cases and 84 controls, so as to draw the ROC curve and evaluate The sensitivity and specificity of the prediction, and then evaluate the ability of detecting the level of this metabolic small molecule in semen to evaluate azoospermia.

[0111] See figure 1 , the sensitivity of 1,3-dihydroxypropan-2-ylpentadecanoate is 96.3%, the specificity is 91.4%, and the area under the ROC curve is 0.965, so 1,3-dihydroxypropan-2-ylpentadecanoate Alkanoates have a better ability to diagnose azoospermia.

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Abstract

The invention relates to a molecular marker related to azoospermia and a detection method and application thereof, and belongs to the technical field of analytical chemistry and clinical medicine. Metabolic small molecules in normal control and azoospermia semen are compared by adopting UPLC-Qexactive MS (Ultra Performance Liquid Chromatography-Quantitative Active Mass Spectrometry), 1, 3-dihydroxypropyl-2-yl pentadecanoate which can be used for evaluating whether the semen suffers from azoospermia or not and has diagnostic value exists in the semen, and the application of the UPLC-Qexactive MS for detecting the 1, 3-dihydroxypropyl-2-yl pentadecanoate is found, so that the 1, 3-dihydroxypropyl-2-yl pentadecanoate can be used for evaluating whether the semen suffers from azoospermia. And the azoospermia diagnosis and monitoring kit convenient for clinical application is developed. Metabolic small molecules are novel biomarkers, are highly associated with disease outcomes, and are stable, non-invasive, easy to detect and accurate in quantification. The 1, 3-dihydroxypropyl-2-yl pentadecanoate provided by the invention can be used as a diagnostic marker of azoospermia, provides a basis for further deep examination of clinicians, provides support for quickly and accurately mastering the disease state and the severity of the disease state of a patient and timely taking a prevention and treatment scheme, and delays and prevents disease progression.

Description

technical field [0001] The invention relates to a molecular marker related to azoospermia and its detection method and application, belonging to the technical fields of analytical chemistry and clinical medicine. Background technique [0002] In 1988, the World Health Organization (WHO) first proposed the concept of reproductive health, that is, the complete and harmonious physiological, psychological and social conditions of human beings and their individuals in the structure, function and behavior of reproductive-related organisms throughout life. health status. Infertility is a common problem among couples worldwide, an estimated 4-17% of couples visit a doctor to correct their infertility, but it is generally believed that more infertile couples do not go diagnosis and treatment. Surveys show that the male factor is the main or only factor accounting for about 20-50% of the causes of infertility. The incidence of azoospermia accounts for 10%-20% of male infertility. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/54G01N30/72C07C69/675
CPCG01N30/02G01N30/54G01N30/72C07C69/675
Inventor 沙家豪袁艳陈敏健杨晓玉王嘉琛姚莹
Owner 苏州南医大创新中心
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